Stable isotope switching (SIS): a new stable isotope probing (SIP) approach to determine carbon flow in the soil food web and dynamics in organic matter pools

RATIONALE: Recent advances in stable isotope probing (SIP) have allowed direct linkage of microbial population structure and function. This paper details a new development of SIP, Stable Isotope Switching (SIS), which allows the simultaneous assessment of carbon (C) uptake, turnover and decay, and the elucidation of soil food webs within complex soils or sedimentary matrices. METHODS: SIS utilises a stable isotope labelling approach whereby the 13C-labelled substrate is switched part way through the incubation to a natural abundance substrate. A 13CH4 SIS study of landfill cover soils from Odcombe (Somerset, UK) was conducted. Carbon assimilation and dissimilation processes were monitored through bulk elemental analysis isotope ratio mass spectrometry and compound-specific gas chromatography/combustion/isotope ratio mass spectrometry, targeting a wide range of biomolecular components including: lipids, proteins and carbohydrates. RESULTS: Carbon assimilation by primary consumers (methanotrophs) and sequential assimilation into secondary (Gram-negative and -positive bacteria) and tertiary consumers (Eukaryotes) was observed. Up to 45% of the bacterial membrane lipid C was determined to be directly derived from CH4 and at the conclusion of the experiment ca. 50% of the bulk soil C derived directly from CH4 was retained within the soil. CONCLUSIONS: This is the first estimate of soil organic carbon derived from CH4 and it is comparable with levels observed in lakes that have high levels of benthic methanogenesis. SIS opens the way for a new generation of SIP studies aimed at elucidating total C dynamics (incorporation, turnover and decay) at the molecular level in a wide range of complex environmental and biological matrices.